Immunohistochemistry was performed by using the peroxidase protocol. Briefly, tissue sections were deparaffinized in xylene, hydrated through descending ethanol, and endogenous peroxidase activity was eliminated by incubation in 3% hydrogen peroxide in methanol for 5 min. After antigen-retrieval in boiling sodium citrate buffer (10 mM), the sections were incubated with primary antibodies for overnight at 4 °C. The signal was developed using Histostain-SP kit (Invitrogen). For the double immunofluorescence staining, the sections were incubated overnight at 4 °C with a mixture of antibodies. After being washed with TBS, the sections were incubated with a mixture of Alexa Fluor 488- and 568-coupled secondary antibodies (Invitrogen) for detection. Images were acquired through Confocal (Olympus FV1000).
Alexa fluor 488 and 568 coupled secondary antibodies
Manufactured by Thermo Fisher Scientific
Alexa Fluor 488- and 568-coupled secondary antibodies are fluorescent dye-labeled antibodies used in various immunodetection techniques. The Alexa Fluor 488 dye emits green fluorescence, while the Alexa Fluor 568 dye emits orange-red fluorescence. These antibodies can be used to detect and visualize target proteins in samples, such as cells or tissue sections.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using alexa fluor 488 and 568 coupled secondary antibodies
1
Immunohistochemistry and Immunofluorescence Staining
2
Immunohistochemistry and Immunofluorescence Staining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
IHC was performed by using the peroxidase protocol. In brief, tissue sections were deparaffinized in xylene, hydrated through descending ethanol, and endogenous peroxidase activity was eliminated by incubation in 3% hydrogen peroxide in methanol for 5 min. After antigen-retrieval in boiling sodium citrate buffer (10 mM), the sections were incubated with primary antibodies for overnight at 4°C. The signal was developed using Histostain-SP kit (Invitrogen). For the double immunofluorescence staining, the sections were incubated overnight at 4°C with a mixture of antibodies. After being washed with TBS, the sections were incubated with a mixture of Alexa Fluor 488– and 568–coupled secondary antibodies (Invitrogen) for detection. Images were acquired through Confocal (Olympus FV1000). The Olympus FV1000 is equipped with four 405-, 488-, 515-, 543-, and 635-nm laser lines and 10× 0.4 numerical aperture (NA), 20× 0.75 NA, 40× 1.3 NA, oil, 60× 1.49 NA, oil, 100× 1.45 NA, and oil objectives. Acquisition Software is Olympus Fluoview v4.2. ImageJ was used for the intensity and colocalization analysis.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!